LOCUS       sequence-304585-       14234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hu6 HGPS sgRNA expression and ABE7.10max VRQR lentiviral plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14234)
  AUTHORS   Koblan LW, Erdos MR, Wilson C, Cabral WA, Levy JM, Xiong ZM, Tavarez
            UL, Davison LM, Gete YG, Mao X, Newby GA, Doherty SP, Narisu N, 
            Sheng Q, Krilow C, Lin CY, Gordon LB, Cao K, Collins FS, Brown JD, 
            Liu DR
  TITLE     In vivo base editing rescues Hutchinson-Gilford progeria syndrome in
            mice.
  JOURNAL   Nature. 2021 Jan;589(7843):608-614. doi: 10.1038/s41586-020-03086-7.
            Epub 2021 Jan 6.
  PUBMED    33408413
REFERENCE   2  (bases 1 to 14234)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41586-020-03086-7"; journalName: "Nature"; date: 
            "2021-01"; volume: "589"; issue: "7843"; pages: "608-614"
COMMENT     Sequence Label: hu6 HGPS sgRNA expression and ABE7.10max VRQR
            ...
FEATURES             Location/Qualifiers
     source          1..14234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2371..2619
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2371..2391
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2542..2561
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2640..2715
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2777..2988
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3038..3058
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4247..8347
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8390..8410
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8420..8476
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8477..9073
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     8970..8990
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9089..9677
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9142..9162)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9560..9571)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9749..9982
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10008..10025)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10014..10238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10284..10712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10371..10390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10581..10602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10721..10741)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10726..11055
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10906..11041
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10968..10987
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11103..11150
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11169..11543
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    11673..11794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11710..11729)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11764..11783
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11843..11859)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11843..11859)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11856..11878)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11867..11883
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(11891..11897,11898..11915,11916..11921))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11936..11957
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12074..12091)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12245..12833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12325..12344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13004..13795,13796..13864))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     13627..13646
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13865..13969)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14044..14063)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat
     2401 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     2461 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     2521 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     2581 ttcttggctt tatatatctt gtggaaagga cgaaacaccg gtccacccac ctgggctccg
     2641 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     2701 gcaccgagtc ggtgcttttt tgaattcgct agctaggtct tgaaaggagt gggaattggc
     2761 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     2821 ggggtcggca attgatccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     2881 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     2941 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagga ccggtgccac
     3001 catgaaacgg acagccgacg gaagcgagtt cgagtcacca aagaagaagc ggaaagtctc
     3061 tgaagtcgag tttagccacg agtattggat gaggcacgca ctgaccctgg caaagcgagc
     3121 atgggatgaa agagaagtcc ccgtgggcgc cgtgctggtg cacaacaata gagtgatcgg
     3181 agagggatgg aacaggccaa tcggccgcca cgaccctacc gcacacgcag agatcatggc
     3241 actgaggcag ggaggcctgg tcatgcagaa ttaccgcctg atcgatgcca ccctgtatgt
     3301 gacactggag ccatgcgtga tgtgcgcagg agcaatgatc cacagcagga tcggaagagt
     3361 ggtgttcgga gcacgggacg ccaagaccgg cgcagcaggc tccctgatgg atgtgctgca
     3421 ccaccccggc atgaaccacc gggtggagat cacagaggga atcctggcag acgagtgcgc
     3481 cgccctgctg agcgatttct ttagaatgcg gagacaggag atcaaggccc agaagaaggc
     3541 acagagctcc accgactctg gaggatctag cggaggatcc tctggaagcg agacaccagg
     3601 cacaagcgag tccgccacac cagagagctc cggcggctcc tccggaggat cctctgaggt
     3661 ggagttttcc cacgagtact ggatgagaca tgccctgacc ctggccaaga gggcacgcga
     3721 tgagagggag gtgcctgtgg gagccgtgct ggtgctgaac aatagagtga tcggcgaggg
     3781 ctggaacaga gccatcggcc tgcacgaccc aacagcccat gccgaaatta tggccctgag
     3841 acagggcggc ctggtcatgc agaactacag actgattgac gccaccctgt acgtgacatt
     3901 cgagccttgc gtgatgtgcg ccggcgccat gatccactct aggatcggcc gcgtggtgtt
     3961 tggcgtgagg aacgcaaaaa ccggcgccgc aggctccctg atggacgtgc tgcactaccc
     4021 cggcatgaat caccgcgtcg aaattaccga gggaatcctg gcagatgaat gtgccgccct
     4081 gctgtgctat ttctttcgga tgcctagaca ggtgttcaat gctcagaaga aggcccagag
     4141 ctccaccgac tccggaggat ctagcggagg ctcctctggc tctgagacac ctggcacaag
     4201 cgagagcgca acacctgaaa gcagcggggg cagcagcggg gggtcagaca agaagtacag
     4261 catcggcctg gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     4321 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     4381 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
     4441 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     4501 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     4561 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     4621 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     4681 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     4741 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     4801 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     4861 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     4921 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     4981 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     5041 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     5101 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     5161 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     5221 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     5281 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     5341 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     5401 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     5461 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     5521 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     5581 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     5641 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     5701 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     5761 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     5821 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     5881 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     5941 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     6001 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     6061 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     6121 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     6181 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     6241 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     6301 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     6361 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     6421 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     6481 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     6541 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     6601 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     6661 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     6721 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     6781 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     6841 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     6901 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     6961 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     7021 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     7081 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     7141 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     7201 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     7261 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     7321 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     7381 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     7441 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     7501 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     7561 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     7621 ggaccctaag aagtacggcg gcttcgtgag ccccaccgtg gcctattctg tgctggtggt
     7681 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     7741 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     7801 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     7861 ggaaaacggc cggaagagaa tgctggcctc agccagagaa ctgcagaagg gaaacgaact
     7921 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     7981 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     8041 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     8101 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     8161 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     8221 ctttgacacc accatcgacc ggaagcagta cagaagcacc aaagaggtgc tggacgccac
     8281 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     8341 aggtgactct ggcggctcaa aaagaaccgc cgacggcagc gaattcgagc ccaagaagaa
     8401 gaggaaagtc ggatccgggg ccaccaactt ttctctgctg aaacaagccg gagatgtcga
     8461 agagaatcct ggaccgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     8521 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     8581 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     8641 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     8701 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     8761 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     8821 ccggcccaag gagcccgcgt ggttcctggc caccgtcgga gtctcgcccg accaccaggg
     8881 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     8941 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     9001 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     9061 gcccggtgcc tgaacgcgtt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     9121 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     9181 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     9241 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     9301 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     9361 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     9421 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     9481 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     9541 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     9601 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     9661 ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt
     9721 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     9781 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     9841 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     9901 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     9961 tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
    10021 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
    10081 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
    10141 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
    10201 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
    10261 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
    10321 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
    10381 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
    10441 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
    10501 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
    10561 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
    10621 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
    10681 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
    10741 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
    10801 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    10861 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
    10921 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
    10981 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
    11041 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
    11101 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
    11161 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
    11221 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
    11281 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
    11341 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
    11401 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
    11461 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
    11521 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
    11581 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
    11641 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
    11701 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
    11761 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
    11821 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
    11881 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
    11941 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
    12001 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
    12061 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
    12121 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
    12181 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
    12241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
    12301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
    12361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
    12421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
    12481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
    12541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
    12601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
    12661 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
    12721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
    12781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
    12841 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
    12901 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
    12961 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
    13021 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
    13081 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
    13141 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
    13201 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
    13261 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
    13321 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
    13381 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
    13441 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
    13501 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
    13561 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    13621 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    13681 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    13741 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    13801 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    13861 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    13921 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    13981 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
    14041 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
    14101 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
    14161 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
    14221 cagatatacg cgtt
//
